This study examined the effects of changes in rumen fermentation during the weaning transition on abundance of transporters involved in volatile fatty acid (VFA) absorption or intracellular pH homeostasis. Holstein bull calves (n = 27) were assigned to 1 of 3 treatment groups in a randomized, complete block design: 2 preweaning groups [animals fed milk only (PRE-M) or milk, calf starter, and hay (PRE-S)] and 1 postweaning group (animals fed milk, starter, and hay with a 2-wk weaning transition; POST-S). Calves were euthanized at 42 d of age (PRE-M and PRE-S) or at 63 d of age (POST-S), and rumen epithelium and rumen fluid samples were collected. Rumen fluid was analyzed for VFA concentration, and rumen epithelium was analyzed for the abundance of VFA transporter monocarboxylate transporter isoform 1 (MCT1) and the intracellular pH regulators sodium bicarbonate co-transporter 1 (NBC1) and sodium-proton exchanger 3 (NHE3) protein. Preweaning, total VFA concentrations tended to increase and NBC1 abundance increased with starter intake. Between pre- and postweaning, total VFA concentrations increased but NHE3 protein abundance decreased. In calves, rumen epithelial development during the weaning transition appears to show more pronounced changes in intracellular pH homeostasis than in VFA transport capacity.